Investigating developmental characteristics of biopsied blastocysts stratified by mitochondrial copy numbers using time-lapse monitoring.

BACKGROUND: For in vitro fertilization (IVF), mitochondrial DNA (mtDNA) levels in the trophectodermal (TE) cells of biopsied blastocysts have been suggested to be associated with the cells' developmental potential. However, scholars have reached differing opinions regarding the use of mtDNA levels as a reliable biomarker for predicting IVF outcomes. Therefore, this study aims to assess the association of mitochondrial copy number measured by mitoscore associated with embryonic developmental characteristics and ploidy. METHODS: This retrospective study analyzed the developmental characteristics of embryos and mtDNA levels in biopsied trophectodermal cells. The analysis was carried out using time-lapse monitoring and next-generation sequencing from September 2021 to September 2022. Five hundred and fifteen blastocysts were biopsied from 88 patients undergoing IVF who met the inclusion criteria. Embryonic morphokinetics and morphology were evaluated at 118 h after insemination using all recorded images. Blastocysts with appropriate morphology on day 5 or 6 underwent TE biopsy and preimplantation genetic testing for aneuploidy (PGT-A). Statistical analysis involved generalized estimating equations, Pearson's chi-squared test, Fisher's exact test, and Kruskal-Wallis test, with a significance level set at P < 0.05. RESULTS: To examine differences in embryonic characteristics between blastocysts with low versus high mitoscores, the blastocysts were divided into quartiles based on their mitoscore. Regarding morphokinetic characteristics, no significant differences in most developmental kinetics and observed cleavage dysmorphisms were discovered. However, blastocysts in mitoscore group 1 had a longer time for reaching 3-cell stage after tPNf (t3; median: 14.4 h) than did those in mitoscore group 2 (median: 13.8 h) and a longer second cell cycle (CC2; median: 11.7 h) than did blastocysts in mitoscore groups 2 (median: 11.3 h) and 4 (median: 11.4 h; P < 0.05). Moreover, blastocysts in mitoscore group 4 had a lower euploid rate (22.6%) and a higher aneuploid rate (59.1%) than did those in the other mitoscore groups (39.6-49.3% and 30.3-43.2%; P < 0.05). The rate of whole-chromosomal alterations in mitoscore group 4 (63.4%) was higher than that in mitoscore groups 1 (47.3%) and 2 (40.1%; P < 0.05). A multivariate logistic regression model was used to analyze associations between the mitoscore and euploidy of elective blastocysts. After accounting for factors that could potentially affect the outcome, the mitoscore still exhibited a negative association with the likelihood of euploidy (adjusted OR = 0.581, 95% CI: 0.396-0.854; P = 0.006). CONCLUSIONS: Blastocysts with varying levels of mitochondrial DNA, identified through biopsies, displayed similar characteristics in their early preimplantation development as observed through time-lapse imaging. However, the mitochondrial DNA level determined by the mitoscore can be used as a standalone predictor of euploidy.

Gas exchange and chlorophyll fluorescence responses of Camellia sinensis grown under various cultivations in different seasons.

Sod culture (SC) and conventional agriculture (CA) represent two distinct field management approaches utilized in the cultivation of tea plants in Taiwan. In this study, we employed gas exchange and chlorophyll fluorescence techniques to assess the impact of SC and CA methods on the photosynthetic machinery of Camellia sinensis cv. TTES No.12 (Jhinhsuan) in response to variable light intensities across different seasons. In spring, at photosynthetic photon flux densities (PPFD) ranging from 800 to 2,000 µmol photon m-2 s-1, the net photosynthesis rate (Pn, 10.43 µmol CO2 m-2 s-1), stomatal conductance (Gs, 126.11 mmol H2O m-2 s-1), electron transport rate (ETR, 137.94), and ΔF/Fm' and Fv/Fm (50.37) values for plants grown using SC were comparatively higher than those cultivated under CA. Conversely, the non-photochemical quenching (NPQ) values for SC-grown plants were relatively lower (3.11) compared to those grown under CA at 800 to 2,000 PPFD in spring. Additionally, when tea plants were exposed to PPFD levels below 1,500 µmol photon m- 2 s- 1, there was a concurrent increase in Pn, Gs, ETR, and NPQ. These photosynthetic parameters are crucial for devising models that optimize cultivation practices across varying seasons and specific tillage requirements, and for predicting photosynthetic and respiratory responses of tea plants to seasonally or artificially altered light irradiances. The observed positive impacts of SC on maximum photosynthetic rate (Amax), Fv/Fm, Gs, water-use efficiency (WUE), and ETR suggest that SC is advantageous for enhancing the productivity of tea plants, thereby offering a more adaptable management model for tea gardens.

Performance of preimplantation genetic testing for aneuploidy in IVF cycles for patients with advanced maternal age, repeat implantation failure, and idiopathic recurrent miscarriage.

OBJECTIVE: The primary objective of this study was to investigate whether preimplantation genetic testing for aneuploidy (PGT-A) of blastocysts through array comparative genomic hybridization (aCGH) improves live birth rates (LBR) in IVF cycles for patients with high prevalence of aneuploidy. MATERIALS AND METHODS: This study included 1389 blastocysts with aCGH results derived from 296 PGT-A cycles in IVF patients with advanced maternal age (AMA) (n = 87, group A), those with repeated implantation failure (RIF) (n = 82, group B), those with recurrent miscarriage (RM) (n = 82, group C), and oocyte donors (OD) (n = 45, young age, as a control group). Another 61 AMA patients without PGT-A procedures were used as a control group for group A. Vitrification was performed after blastocyst biopsy, and thawed euploid embryos were transferred in a nonstimulated cycle. RESULTS: For the AMA group, a significant increase in LBRs was found in the PGT-A group compared with the non-PGT-A group (54.1% vs. 32.8%, p = 0.018). Consistent LBRs (54.1%, 51.6%, 55.9%, and 57.1%, respectively, in group A, B, C, and young age group) were obtained for all the indications. CONCLUSIONS: LBRs can be improved using PGT-A of blastocysts with aCGH in IVF cycles for patients with a high rate of aneuploidy, especially for patients with AMA.

Photosynthetic gas exchange responses of Swietenia macrophylla King and Melia azedarach L. plantations under drought conditions.

BACKGROUND: The environmental stresses caused by climate change have become more severe in recent decades, affecting tree growth and physiology. Tropical forests have great potential for global carbon sequestration. However, they suffer from heavy rainfall and prolonged dry periods due to climate change. Swietenia macrophylla King and Melia azedarach L. are economically valuable trees that are widely planted in southern Taiwan. Plantations are exposed to either prolonged dry periods or heavy rainfall within the seasons of tropical monsoon areas. Photo-physiological comparisons may provide information that can improve management of S. macrophylla and M. azedarach plantations in tropical regions. RESULTS: Both species exhibited a midday depression in leaf photosynthesis regardless of the season. The net photosynthetic rate (P N), stomatal conductance (g s), and transpiration rate (E) in the dry season all significantly decreased in both tree species. In addition, M. azedarach used water more efficiently than did S. macrophylla during the dry season, but S. macrophylla had higher P N compared with that in M. azedarach during the wet season. Temperature and vapor pressure deficit influenced P N variation in S. macrophylla and M. azedarach, respectively. Our data suggested that the P N and g s of M. azedarach, but not of S. macrophylla, were linearly correlated during the dry season. The reduction of the leaf area was more sever in M. azedarach than in S. macrophylla, thus preventing water loss more efficiently. CONCLUSIONS: M. azedarach adapted to drought by reducing total leaf area and maintaining higher P N, g s, E, and WUE compared with those measured in S. macrophylla during the dry season. M. azedarach is more drought adaptation and more suitable for both humid and semi-humid areas than S. macrophylla, whereas the latter should be limited to more humid areas.

Requirement of Leukemia Inhibitory Factor or Epidermal Growth Factor for Pre-Implantation Embryogenesis via JAK/STAT3 Signaling Pathways.

Leukemia inhibitory factor (LIF) plays a key role in the survivability of mouse embryos during pre-implantation. In this study, we verified the role of LIF by detecting gene expression in morula stage embryos through DNA microarray. Our results showed that LIF knockdown affected expression of 369 genes. After LIF supplementation, the epidermal growth factor (EGF) is most affected by LIF expression. To observe the correlation between LIF and EGF, the LIF knockdown embryos were supplemented with various growth factors, including LIF, EGF, GM-CSF, TGF, and IGF II. Only LIF and EGF caused the rate of blastocyst development to recover significantly from 52% of control to 83% and 93%, respectively. All of the variables, including the diameter of blastocysts, the number of blastomeres, and cells in ICM and TE, were almost restored. Moreover, EGF knockdown also impaired blastocyst development, which was reversed by LIF or EGF supplementation. The treatment with various signaling suppressors revealed that both EGF and LIF promoted embryonic development through the JAK/STAT3 signaling pathway. These data suggest that the EGF and LIF can be compensatory to each other during early embryonic development, and at least one of them is necessary for sustaining the normal development of pre-implantation embryos.

Leukemia inhibitory factor antisense oligonucleotide inhibits the development of murine embryos at preimplantation stages.

Leukemia inhibitory factor (LIF) is an essential factor for implantation and establishment of pregnancy. However, its role in the development of preimplantation embryos remains controversial. In this study, changes in preimplantation embryos were determined after microinjection of LIF antisense oligonucleotide at the two-pronucleus stage. Although no significant differences were found in the percentages between the untreated group and the 0.25-fmol-treated group, the 0.5- or 1.0-fmol-treated groups had significantly lower percentages of embryos developed to the morula or blastocyst stage and the 2.0-fmol-treated group had significantly lower percentages of embryos developed to the four-cell, morula, or blastocyst stage. No embryos developed to the four-cell stage in the 4.0-fmol-treated group. Moreover, there was a decreasing trend in the levels of LIF immunoactivity with the increasing amount of LIF antisense oligonucleotide injected. The diameter of blastocysts in the 2.0-fmol-treated group was significantly smaller than that in the untreated group. The blastocysts in this group had significantly lower numbers of blastomeres and cells in the inner cell mass (ICM) or trophectoderm (TE) and ICM:TE ratio. The 1.0- or 2.0-fmol-treated groups had significantly lower implantation rates than their corresponding control groups. In the 2.0-fmol groups with supplementing exogenous LIF, significantly lower percentages were also observed in the four-cell, morula, and blastocyst stages. However, blastocysts treated with 50 ng/ml LIF had a significantly higher percentage than those in the LIF gene-impaired group without LIF supplement. These results indicate that LIF is a critical factor for the normal development of embryos at the preimplantation stages.